The effects of 1,25-dihydroxyvitamin D3 on osteoclast formation in fetal mouse metatarsal organ cultures.

We have previously described a model system, using 15-day fetal mouse metatarsals cultured in serumless medium, in which osteoclasts and their precursors develop from in situ progenitors in a manner which is similar, both temporally and spatially, to that which occurs in vivo. In this report we evaluate the role of the osteotropic hormone 1,25-dihydroxyvitamin D3 (1,25-D3) on osteoclast formation in this model system by characterizing its effects on proliferation, differentiation, and fusion of cells of the osteoclast lineage. Morphologic evaluation was used to enumerate osteoclast precursors, mono- and multinucleate osteoclasts, and osteoclast nuclei in serial paraffin sections. Dose response data reveal a significant stimulation of osteoclast formation by 1,25-D3 in the range of 0.6 nM to 40 nM, and kinetic analyses suggest that these effects are on proliferation of osteoclast progenitors as well as on differentiation of precursors to form osteoclasts. A single 48 h exposure between day 4 and 6 of culture is necessary and sufficient to induce maximal osteoclast formation, while continuous exposure beyond this "critical period" inhibits multinucleate osteoclast formation. Simultaneous treatment with indomethacin inhibits the effects of 1,25-D3, while treatment with PGE2 stimulates osteoclast formation without significantly increasing precursor numbers, or inhibiting multinucleate osteoclast formation. These results suggest that the effect of 1,25-D3 to induce differentiation of precursors to form mono- and multinucleate osteoclasts is mediated by endogenous prostaglandin synthesis. On the other hand, the inhibition of polykaryon formation observed with continuous 1,25-D3 treatment, does not appear to be a prostaglandin mediated phenomena.(ABSTRACT TRUNCATED AT 250 WORDS)